EP0097455A2 - Appareil et méthode pour le traitement de fluides dans un champ de force centrifuge - Google Patents

Appareil et méthode pour le traitement de fluides dans un champ de force centrifuge Download PDF

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Publication number
EP0097455A2
EP0097455A2 EP83303334A EP83303334A EP0097455A2 EP 0097455 A2 EP0097455 A2 EP 0097455A2 EP 83303334 A EP83303334 A EP 83303334A EP 83303334 A EP83303334 A EP 83303334A EP 0097455 A2 EP0097455 A2 EP 0097455A2
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EP
European Patent Office
Prior art keywords
bag
component
fluid
port
flexible bag
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Application number
EP83303334A
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German (de)
English (en)
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EP0097455A3 (fr
Inventor
Richard M. Lueptow
Jeffrey J. Peterson
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Haemonetics Corp
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Haemonetics Corp
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Publication date
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Publication of EP0097455A2 publication Critical patent/EP0097455A2/fr
Publication of EP0097455A3 publication Critical patent/EP0097455A3/fr
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
    • B04B—CENTRIFUGES
    • B04B5/00—Other centrifuges
    • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
    • B04B5/0407—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles
    • B04B5/0428—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles with flexible receptacles

Definitions

  • This invention is in the field of fluid processing and more particularly relates to the centrifugal separation of fluid, such as blood, into two or more components, such as during cell washing or component separation.
  • red blood cells red blood cells
  • packed RBC's will hereafter be used to refer to unwashed RBC's which have been separated from other whole blood components.
  • the packed RBC's are washed with a wash solution, such as saline, to remove such undesirable components remaining after the initial centrifugal separation.
  • a wash solution such as saline
  • the packed RBC's can be washed in a number of ways.
  • One method now in practice is to centrifuge a unit of donor blood in a collection bag and subsequently remove the plasma and buffy coat manually using a plasma expressor leaving packed RBC's in the collection bag. Then the packed RBC's are diluted with saline, centrifuged again, and the supernatant manually removed using an expressor, leaving washed RBC's.
  • Packed RBC's can also be washed by diluting the packed RBC's with saline in a centrifugal processing bag or bowl and expressing the supernatant through a rotating seal leaving washed cells.
  • the Haemonetics Model 102 cell washing equipment is of the bowl type. (See: The Preparation of Leukocyte-Poor Red Blood Cells: A Comparative Study Meryman et al., Transfusion 20(3):285:287,1980).
  • the IBM 2991 Cell Washer (generally described in U.S. Patents 4,007,871 and 4,010,894) utilizes a spin and agitation method in which packed RBC's are spun within a saline solution in a toroidal chamber of fixed volume and then agitated in the chamber. This process is repeated many times with fresh wash solution until sufficient hematocrit of the washed RBC's is attained. The agitation is required in order to maximize interaction between the wash solution and the packed RBC's.
  • the IBM 2991 is effective in washing but the apparatus is complex and thus expensive and the procedure very time consuming. (See: Use and Analysis of Saline Washed Red Blood Cells Wooten, M.J., Transfusion 16(5):464 1976).
  • washing apparatus and methods which are simple, inexpensive and speedy.
  • apparatus for processing fluids in a centrifugal force field to separate constituent components of such fluids comprises in combination a centrifuge having a rotor adapted to rotate at a sufficient speed to cause said components to separate, a first flexible bag mounted on the rotor and adapted to contain a first fluid, a receiver container mounted on the rotor and adapted to receive at least one component of said first fluid, a first conduit means for coupling the flexible bag and the receiver container in fluid communication and a first mass means disposed nearer the center of rotation of the rotor than the flexible bag and adapted to move against a surface of said bag, said mass being sufficient to at least initiate a flow from said bag to said container through said conduit means of component fluid separated in said bag and is characterized by a first support means for orienting the flexible bag in the rotor such that an output port on said flexible bag is located nearer the axis of the center of rotation of the centrifuge rotor than an input port on said flexible bag whereby during centri
  • the present invention also includes fluid processing apparatus for separation of fluid components by centrifugation comprising a first flexible bag wherein lighter density fluid components accumulate at a first location within said bag and heavier density fluid components accumulate at a second location within said bag characterized in that said second location is diagonally opposite said first location and ports are located at said first and second location.
  • the present invention still further provides a method for processing fluids in a centrifugal force field to separate constituent components of such fluids comprising mounting a fluid processing chamber on a centrifuge rotor and rotating the rotor and causing a first fluid component to flow through an outlet port of said chamber through a conduit into a receiver container characterized by the step of causing less dense and more dense components of the fluid to accumulate at diagonally opposite corners of said chamber by orienting the chamber with respect to the axis of the centre of rotation of the centrifuge.
  • packed RBC's may be washed by a suitable wash solution within a centrifugal processing bag.
  • the efficiency of the cell washing procedure is optimized by initially orienting the processing bag with respect to the axis of the centrifuge center of rotation (CR), such that (1) the highest density component, i.e., washed RBC's, accumulate at a corner of the bag furthest from the axis of the CR and locating the inlet port for the wash solution at that corner and (2) the lowest density component, i.e., supernatant accumulates at a corner of the bag which is closest to the axis of the CR and locating the outlet port at this latter corner.
  • CR centrifuge center of rotation
  • This orientation may conveniently be accomplished by means of a "double angled" cassette support which forces the processing bag to assume a position in the centrifuge rotor which is at an angle with respect to the axis of rotation and also at an angle with respect to the position of concentricity; hence the term "double angled”.
  • the cassette support member is tilted inwardly from the vertical plane and the cylindrical segment shape of the member is off-set to be eccentric with the axis of the CR.
  • the whole blood bag may be rectangularly shaped with four corners labelled A, B, C and D, counterclockwise from the upper right-hand corner "A" (looking from the CR).
  • the bag is positioned adjacent the double angle support member.
  • the tilted eccentric shape of the support member forces the bag to assume an orientation with respect to the axis of the CR such that: and wherein rl r2, r 4 and r 3 are the radii from respective bag corners A, B, C and D to the axis of the CR.
  • the supernatant outlet port is located at the shortest radius r l , in this case, the upper right-hand corner "A", and the wash solution input port is located at the longest radius r 4 , in this case, the lower left-hand corner "C".
  • the outlet port With the outlet port at the shortest radius, the lower density supernatant component can be readily removed through this port.
  • the wash solution interacts with or "sees” the most packed RBC's. Also, a turbulent flow is created whereby the cells are agitated thereby maximizing the cell washing efficiency.
  • these illustrate an apparatus for performing a method for washing blood components, particularly packed RBC's, with a suitable wash solution, such as a saline solution.
  • a suitable wash solution such as a saline solution.
  • the method and apparatus is not, however, intended to be limited to cell washing and may find applicability in other areas, such as plasma-pheresis or plateletpheresis.
  • the invention may employ a specific centrifuge apparatus found in certain copending applications. Because of the imbalances produced in the processes to be described, it is desirable that a Self-Balancing Centrifuge as described in Application No. 82303592.8 (hereby incorporated by reference), or equivalent, will supply the necessary centrifugal force for fluid processing in accordance with the invention. However, the invention is not intended to be limited to any particular centrifuge.
  • a first container in the form of a flexible bag containing anticoagulated whole blood to be centrifugally separated, is supported by a cassette located on a centrifuge rotor.
  • the cassette is in the form of a rack or stand partitioned into three annular sections by two vertically positioned support members. Each member has a shape generally described by a segment of a hollow cylinder with a radius of curvature corresponding to a radius to the axis of the center of rotation (CR) of the centrifuge rotor.
  • a second container is disposed in the cassette adjacent the first container and in fluid communication with the first container.
  • the second container which may also be a flexible bag, is adapted to receive one or more of the centrifugally separated components of the anticoagulated whole blood.
  • a pressure plate in the form of a body of material such as a metal plate also having a curvature corresponding to a radius to the axis of the centrifuge CR, and having a predetermined mass is disposed between the first bag and the center of rotation of the rotor.
  • This pressure plate is suspended so that it is free to move radially against the first bag when subjected to the centrifugal forces generated by rotation of the centrifuge.
  • the pressure plate has a predetermined mass sufficient to at least initiate a flow of separated fluid component from the first bag to the second bag as the pressure plate presses against the first bag during rotation of the centrifuge rotor.
  • the mass distribution and shape of the pressure plate is adapted to pool the separated first blood component in the area of an umbilical fitment on the bag. An output port is located at this fitment.
  • the first bag and second bag are located adjacent each other on the rotor with the first bag positioned radially inward from the second bag.
  • a siphon effect is created when flow is initiated from the first bag to the second bag as the pressure plate pushes against the first bag under the influence of centrifugal force.
  • the siphon effect is due to the difference in centrifugal forces to which the bags are subjected because one bag is located nearer the center of rotation than the other.
  • a double-angle cassette support member 9 (to be described in detail in connection with Figs. 6 and 7) is located on one side of a centrifuge rotor 28 near the periphery.
  • a weight plate 15 is located adjacent the cassette support member 9 and is free to move radially under the influence of centrifugal force toward the support member 9.
  • a blood processing/ cell washing bag 8 is disposed between the weight plate 15 and the support member 9 and is oriented by support member 9 in a position such that lighter density component accumulates at the upper right-hand corner "A" of the bag 8 and heavier density component accumulates at the lower left-hand corner "C” of the bag 8.
  • a deformable support member 100 is provided at corner "A" to ensure that the outlet at port "A" is located sufficiently near the axis of the CR to enable all of the lighter density component (supernatant) to exit the port located at corner "A".
  • the bag 8 is in fluid communication with (1) the wash solution bag 20 via wash line 25, (2) a supernatant bag 2 via supernatant line 27, and (3) a packed RBC's bag 4 via fill line 23.
  • Line 27 is coupled through a solenoid actuated clamp 92c.
  • Wash line 25 is coupled through a second solenoid actuated clamp 92d.
  • Each of these clamps are supported on vertical support members 74a and 74b, which along with vertical support member 74c form an H-shaped vertical support structure to which various fixed components of the cell washing process may be attached.
  • Supernatant bag 2 is disposed at the periphery of the centrifuge rotor opposite the double-angled cassette support member 9.
  • Wash solution bag 20 is located between a cassette support 11 and a weight plate 17 at a location nearer the axis of the CR of the rotor than the blood processing/cell washing bag 8, but on the opposite side of the axis of the CR from bag 8.
  • the packed RBC's bag 4 is located between a RBC cassette support 13 and a weight plate 19 at a location nearer the axis of the CR of the rotor than the blood processing bag 8 and on the same side of the CR as bag 8.
  • the bag 4 is placed in the RBC cassette between the support 13 and the weight plate 19.
  • the packed RBC's bag 4 is connected by bag spike 43a (see Fig. 3) and conduit 23 to processing bag 8.
  • Processing bag 8 is placed between the double angle cassette 9 and weight plate 15.
  • the wash solution bag 20 is attached to the processing bag 8 with bag spike 43b (see Fig. 3) and placed between cassette support 11 and weight plate 17. All of the weight plates and supports, with the exception of the support member 9 and weight plate 15, are similar to those described in the previously referenced Application No. 82303594.4.
  • the conduit from supernatant bag 2 is labelled 27 in Fig. 3, and as seen in Figs. 1, 2 and 5 is disposed between optical sensor 90 and solenoid actuator clamp 92c.
  • the conduit 25 between the port 47 (Fig. 3) and the wash solution bag 20 is disposed between a solenoid actuated clamp 92d (see Fig. 1).
  • the control circuitry for the clamps 92c and 92d is shown in Fig. 5.
  • the anparatus is now ready for a cell washing procedure.
  • the centrifuge is rotated, causing pressure plate 19 to press against packed RBC's bag 4 expressing the contents into the processing bag 8.
  • conduit 27 has been clamped off by clamp 92c.
  • conduit 25 is clamped until sufficient dwell time is achieved to ensure that the RBC's have accumulated at port 47.
  • the clamp 92d on conduit 25 is opened, the wash solution is expressed into the processing bag 8, now containing packed RBC's. This is accomplished while the centrifuge is spinning and cell washing takes place, as shown generally in Fig. 4.
  • conduit 27 connected to port 45 of the processing bag is unclamped by operation of clamp 92c to permit passage of supernatant from processing bag 8 to supernatant bag 2.
  • a siphon effect is created when flow is initiated from the processing bag 8 to the supernatant bag 2 as the pressure plate 15 pushes against the processing bag under the influence of centrifugal force.
  • the siphon effect is due to the difference in centrifugal forces to which the bags are subjected because one bag is located nearer the center of rotation than the other.
  • Optical sensor 90 senses when red blood cells pass through conduit 27 whereupon it provides a signal to control 94a (see Fig. 5) which energizes clamp 92c to clamp conduit 27 and prevents further flow from the processing bag 8.
  • the procedure of expressing saline into the processing bag 8 through conduit 25 and removing supernatant from processing bag 8 through conduit 27 may be repeated several times to assure an optimal removal of plasma, platelets, white blood cells and cell debris from the packed RBC's.
  • the centrifuge can be stopped, and the conduit 27 to the supernatant bag 2 may be manually clamped and severed from the processing bag 8 which now contains the washed RBC's.
  • the packed RBC's bag 4 and the wash bag 20 may be severed from the processing bag 8 and the RBC'S, which have now been centrifugally washed, may be reintroduced to a patient.
  • the processing bag 8 is oriented by a rigid back support member 9, such that (1) the highest density component (RBC's) accumulate at the lower left-hand corner of the bag furthest from the axis of the center of rotation (CR) of the centrifuge and where the inlet port 47 is located and (2) the lowest density component (supernatant) and wash solution accumulate at the upper right-hand corner of the bag which is closest to the axis of the CR of the centrifuge and where the outlet port 45 is located, as shown in Fig. 7.
  • RBC's highest density component
  • CR center of rotation
  • the cassette support member 9 is tilted inwardly from the vertical plane and the hollow cylindrical segment shape of the member 9 is formed eccentric with the axis of the CR.
  • the degree of tilt is preferably sufficient to provide a maximum separation gradient consistent with the permissible space provided in the centrifuge rotor.
  • the degree of eccentricity of the support member is predicted on achieving good separation within the limitations of space.
  • the pressure plate (15 in Fig. 7) is a body of material such as a metal or plastics plate having a curvature generally corresponding to the curvature of the support member 9 and having a predetermined mass.
  • the plate is disposed between the processing bag and the axis of the CR of the rotor.
  • This pressure plate 15 is suspended so that it is free to move radially against the processing bag 8 when subjected to the centrifugal forces generated by rotation of the centrifuge.
  • the pressure plate 15 has a predetermined mass sufficient to at least initiate a flow of separated fluid component from the processing bag 8 to the supernatant bag 2 as the pressure plate 15 presses against the processing bag 8 during rotation of the centrifuge rotor.
  • the mass distribution and shape of the pressure plate 15 is adapted to pool the separated component in the area of the outlet port 45.
  • Bag 8 is rectangular in shape (as can be seen more clearly in Fig. 4 with four corners A, B, C, and D, lettered counterclockwise from the upper right-hand corner.
  • the bag is manufactured from two sheets of PVC welded together at the edges.
  • the tilted eccentric shape of the support member forces the bag to assume an orientation, with respect to the axis of the CR (see Fig. 6) such that and wherein r l , r 2 , r 4 and r 3 are the radii from respective bag corners A, B, C and D to the axis of the CR.
  • the outlet port 45 to the supernatant bag 2 is located at the shortest radius r 1 , in this case, the upper right-hand corner "A", and the wash solution input port 47 is located at the longest radius r 4 , in this case the lower left-hand corner "C".
  • the outlet port With the outlet port at the shortest radius, the lower density component can be removed through this port. Furthermore, by introducing the wash solution at the longest radius port location, the solution "sees" the most packed RBC's and generates a counter-current flow through the red cells, thus maximizing cell washing efficiency.
  • the software set consists essentially of two fluid interconnected flexible bags 2 and 8, plus two accessory bags 4 and 20. Bags 4 and 20 are not initially interconnected with the other bags but bag 8 is equipped with conduits 23 and 25, respectively, at the end of which bag spikes 43b and 43a are provided to enable fluid communication with ports 42 on each bag 4 and 20 respectively for cell washing.
  • Bag 4 contains packed RBC's which are expressed into processing bag 8 via port 40 for cell washing with a wash solution from bag 20.
  • the wash solution is expressed into corner port 47 from bag 20 via conduit 25.
  • Supernatant from the wash procedure is expressed from bag 8 via outlet corner port 45 and conduit 27 to supernatant bag 2.
  • these bags are preferably made of suitable thin walled hemo-compatible plastics material, such as polyvinyl chloride (PVC).
  • PVC polyvinyl chloride
  • the saline carries other less dense components (supernatant) such as platelets, white blood cells and plasma proteins with it leaving only washed red cells in corner "C".
  • supernatant including the platelets, white blood cells and plasma proteins can then be expressed through port 45 at corner "A".
  • the wash inlet port 47 is small enough in diameter to provide a turbulent jet to promote mixing of the wash solution and RBC's.
  • a 1/16 inch diameter inlet port with a 300 ml/minute saline wash solution flow rate provides acceptable results.
  • the inlet port 47 at "C” works best if it is disposed at an angle of from 30° to 90° with respect to the side of the bag. This directs the saline into the center of where the RBC's have packed. It may also be desirable to create a more diagonal shaped bag as indicated by the dashed lines in Fig. 4. This would prevent packing in corners "B" and "D"'where the wash solution jet may not reach the cells.
  • the radius r 4 at "C” was 5.1 inches
  • the radius r 3 at “D” was 5.1 inches
  • the radius r 5 at “E” was 5.5 inches. Consequently, the red blood cells accumulated at "E” instead of "C” where they were desired.
  • an increasing radius may be desirable for certain applications. For example, if it were desired to retain some plasma with packed red cells to lower the hematocrit, a pocket of plasma could be retained by first increasing the radius from the bottom of the bag (C and D) to midway up the bag and then decreasing the radius from the midpoint to the top of the bag (A and B).
  • the outlet port 45 must be positioned at an innermost radius as shown in Fig. 8A. If the port is allowed to move out towards the back support member due to the centrifugal force, some components may be trapped at an inner radius, as happened to component A in Fig. 8B.
  • the outlet port may be held directly to the weight plate 15 with clips, or a deformable support member 100 may be used to position the outlet port 45 against the weight plate.
  • This deformable support member 100 may be a spring mechanism or (as shown) may be shaped from a deformable material such as foam rubber.
  • a measure of the effectiveness of a wash procedure with respect to the removal of plasma proteins is the fraction of free hemoglobin removed as a function of the amount of wash solution used.
  • the hemoglobin content of the fluid surrounding the the RBC's is easily measured. Determining the hemoglobin content before and after a wash procedure provides a quantitative measure of the fraction of plasma proteins removed.
  • the IBM 2991 cell washing system removes 99.4% of the plasma (as measured by total protein) and consumes 1000 ml of saline (M.J. O'Connor Wooten, Transfusion 16(5): 464-468 (1976)) and takes about 27 minutes (H.T. Meryman, et al.., Transfusion 20(3): 285-292(1980)).
  • the processing bag 8 may be used purely for pheresis (component separation) in which case, anticoagulated whole blood may be introduced at the lower corner "C" (port 47) and centrifugally separated into packed RBC's and plasma. After separation, the plasma would be expressed out corner "A" (port 45) in the manner previously described.
  • the apparatus may be used for deglycerolization of frozen RBC's in glycerol.
  • the frozen product is thawed and introduced into bag 8 at corner "B" (port 40) and processed as previously described until the glycerol is removed with the supernatant.
  • support member 9 and pressure plate 15 may be described in general as segments of a hollow cylinder, they need not be cylindrically shaped but can be asymetric in shape to provide pooling of components at desired locations.
EP83303334A 1982-06-17 1983-06-09 Appareil et méthode pour le traitement de fluides dans un champ de force centrifuge Withdrawn EP0097455A3 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US06/389,242 US4482342A (en) 1982-06-17 1982-06-17 Blood processing system for cell washing
US389242 1982-06-17

Publications (2)

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EP0097455A2 true EP0097455A2 (fr) 1984-01-04
EP0097455A3 EP0097455A3 (fr) 1985-07-24

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EP (1) EP0097455A3 (fr)
JP (1) JPS596953A (fr)
AU (1) AU1319483A (fr)
DK (1) DK278583A (fr)

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DK278583A (da) 1983-12-18
AU1319483A (en) 1983-12-22
US4482342A (en) 1984-11-13
JPS596953A (ja) 1984-01-14
EP0097455A3 (fr) 1985-07-24
DK278583D0 (da) 1983-06-16

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